血液・血管システム形成の分子制御プログラム

2004 Fiscal Year Annual Research Report

• Project/Area Number: 15390086
• Research Institution: University of Tsukuba
• Principal Investigator: 山本 雅之 筑波大学, 大学院・人間総合科学研究科, 教授 (50166823)
• Co-Investigator(Kenkyū-buntansha): 清水 律子 筑波大学, 大学院・人間総合科学研究科, 講師 (40226262)
• Keywords: 造血幹細胞 / GATA-2 / GFPノックインマウス / 条件付きノックアウトマウス

Research Abstract

GATA-2遺伝子の血球特異的なプロモーター・第1エクソンの下流に緑色蛍光蛋白質(GFP)遺伝子を挿入したマウス(ノックインノックアウトマウス:G2-GFP)を樹立し,骨髄におけるGATA-2発現細胞の同定および解析を行なった.ヘテロマウスの骨髄では、全細胞の約0.5%の割合でGFP陽性細胞が存在し,これらの細胞は血球分化マーカー陰性(Lin^-),c-Kit^+の造血前駆細胞およびLin^-/c-Kit^+/Sca1^+の造血幹細胞であった.GFP陽性細胞をOP9ストローマ細胞と共培養したところ,30日以上にわたり多系統の血球細胞を産生したことから,この細胞は未分化で増殖能の高い細胞であることが示唆された.このGFP陽性細胞では,c-Myc,Cyclin A2,Cyclin B1等の増殖中の細胞で発現する遺伝子が,GFP陰性細胞に比べて多く発現していた.さらに,Lin^-/c-Kit^+/GFP^+分画には,30%以上の高い割合でS/G2/M期の細胞が含まれていた.これらの結果から,GATA-2は骨髄造血幹細胞および前駆細胞の細胞周期進行に関与する遺伝子発現を制御していることが示唆される.
このようなGATA-2の造血前駆細胞の増殖における機能をより詳細に解析するために,loxP配列を両端に有するGATA-2 cDNAをGATA-2遺伝子の翻訳開始部位にノックインする条件付きノックアウトマウスの作成した(G2-CKO).GATA-2ノックアウトマウスは胎生致死となるため,本マウスによってのみ成獣骨髄でのGATA-2の機能を検討することができる.すでに,同マウスをインターフェロン投与により誘導的にCreリコンビナーゼを発現するMxl-Creトランスジェニックマウス,および,上述のGFPノックインマウスと交配し,三重複合体(G2^<GFP/CKO>::Mx1-Cre)を確立している.現在,骨髄移植法を用いた骨髄造血前駆細胞におけるGATA-2の機能関与をGFPを指標として検討する系を立ち上げている.

Research Products

• [Journal Article] Transgenic overexpression of GATA-1 mutant lacking N-finger domain causes hemolytic syndrome in mouse erythroid cells. (2005)
  • Author(s): Nakano, M., Ohneda, K., Mukai, H.Y., Shimizu, R., Ohneda, O., Ohmura, S., Suzuki, M., Tsukamoto, S., Yanagawa, T., Yoshida, H., Takakuwa, Y, Yamamoto, M.
  • Journal Title: Genes Cells 10 Pages: 47-62
• [Journal Article] Nrf2 transcriptionally activates the mafG gene through an antioxidant response element. (2005)
  • Author(s): Katsuoka, F., Motohashi, M., Engel, J.D., Yamamoto, M.
  • Journal Title: J.Biol.Chem. 280 Pages: 4483-4490
• [Journal Article] Evolutionary conserved N-terminal domain of Nrf2 is essential for the Keapl-mediated degradation of the protein by proteasome. (2005)
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  • Journal Title: Arch.Biochem.Biophys. 433 Pages: 342-350
• [Journal Article] Purification, crystallization and preliminary X-ray diffraction analysis of the Kelch-like motif region of mouse Keap1. (2005)
  • Author(s): Padmanabhan, B., Scharlock, M, Tong, K.I, Nakamura, Y., Kang, M-I., Kobayashi, A., Matsumoto, T., Tanaka, A., Yamamoto, M, Yokoyama, S.
  • Journal Title: Acta Crystallographica 61 Pages: 153-155
• [Journal Article] Essential role of synoviolin in embryogenesis. (2005)
  • Author(s): Yagishita, N., Ohneda, K., Amano, T., Yamasaki, S., Sugiura, A., Tsuchimochi, K., Shin, H., Kawahara, K., Ohneda, O., Ohta, T., Tanaka, S., Yamamoto, M., Maruyama, I., Nishioka, K., Fukamizu, A., Nakajima, T.
  • Journal Title: J.Biol.Chem. (印刷中)
• [Journal Article] Restricted expression of constitutively active-aryl hydrocarbon receptor in T lineage cells causes thymus involution and suppresses immunization-induced splenocyte increase. (2005)
  • Author(s): Nohara, K, Pan, X., Tsukumo, S., Hida, A., Ito, T., Nagai, H., Inouye, K., Motohashi, H., Yamamoto, M., Fujii-Kudyama, Y, Tohyama, C.
  • Journal Title: J.Immunol (印刷中)
• [Journal Article] Pi-class glutathione S-transferase genes are regulated by Nrf2 through an evolutionarily conserved regulatory element in zebrafish. (2005)
  • Author(s): Suzuki, T., Takagi, Y., Osanai, H., Li, L., Takeuchi, M., Katoh, Y., Kobayashi, M, Yamamoto, M
  • Journal Title: Biochem.J. (印刷中)
• [Journal Article] UVA irradiation induces Nrf2 activation in dermal fibroblasts : Protective role in UVA-induced apoptosis. (2005)
  • Author(s): Hirota, A., Kawachi, Y., Itoh, K., Nakamura, Y., Xu, X., Banno, T., Takahashi, T., Yamamoto, M, Ohtsuka, F.
  • Journal Title: Invest.Dermatol. (印刷中)
• [Journal Article] The distal sequence element of the selenocysteine tRNA gene is a tissue dependant enhancer essential for mouse embryogenesis. (2005)
  • Author(s): Kelly, V.P., Suzuki, T., Nakajima, O., Arai, T., Tamai, Y., Takahashi, S., Nishimura.S, Yamamoto, M.
  • Journal Title: Mol.Cell.Biol. (印刷中)
• [Journal Article] GATA transcription factors inhibit cytokine-dependent growth and survival of a hematopoietic cell line through the inhibition of STAT3 activity. (2005)
  • Author(s): Ezoe, S., Matsumura, I., Gale, K, Satoh, Y., Ishikawa, J., Mizuki, M., Takahashi, S., Minegishi, N., Nakajima, K, Yamamoto, M., Enver, T, Kanakura, Y.
  • Journal Title: J.Biol.Chem.
• [Journal Article] Role of 15-deoxyD^<12-14> Prostaglandin J_2 and Nrf2 Pathways in Protection against Acute Lung Injury. (2005)
  • Author(s): Mochizuki, M., Ishii, Y., Itoh, K., Iizuka, T., Morishima, Y., Kimura, T., Kiwamoto, T., Matsuno, Y., Hegab, A.E., Nomura, A., Sakamoto, T., Uchida, K., Yamamoto, M, Sekizawa, K.
  • Journal Title: Am.J.Res.Cri.Care Med (印刷中)
• [Journal Article] Molecular mechanisms activating Nrf2-Keap1 pathway in regulation of antioxidant gene. (2005)
  • Author(s): Kobayashi, M, Yamamoto, M.
  • Journal Title: Antioxidants Redox Signaling 7 Pages: 384-395
• [Journal Article] Gene expression regulation and domain function of hematopoietic GATA factors. (2005)
  • Author(s): Shimizu, R, Yamamoto, M.
  • Journal Title: Seminars in Cell & Developmental Biology 16 Pages: 129-136
• [Journal Article] GATA1 function, a paradigm for transcription factors in hematopoiesis. (2005)
  • Author(s): Ferreira, R., Ohneda, K., Yamamoto, M., Philipsen, S.
  • Journal Title: Mol.Cell.Biol. 25 Pages: 1215-1217
• [Journal Article] Transgenic rescue of GATA-1-deficient mice with GATA-1 lacking a FOG-1 association site phenocopies patients with X-linked thrombocytopenia. (2004)
  • Author(s): Shimizu, R., Ohneda, K., Engel, J.D., Trainor, C.D., Yamamoto, M.
  • Journal Title: Blood 103 Pages: 2560-2567
• [Journal Article] Role of Nrf2 in the regulation of CD36 and stress protein expression in murine macrophages : activation by oxidatively modified LDL and 4-hydroxynonenal (2004)
  • Author(s): Ishii, T., Itoh, K., Ruiz, E., Leake, D.S., Unoki, H., Yamamoto, M., Mann, G.E.
  • Journal Title: Circulation 94 Pages: 609-616
• [Journal Article] Protection against electrophile and oxidant stress by induction of the phase 2 response : Fate of cysteines of the Keap1 sensor modified by inducers. (2004)
  • Author(s): Wakabayashi, N., Dinkova-Kostova, A.T.Holizclaw, W.D., Kang, M-L, Kobayashi, A., Yamamoto, M., Kensler, T.W., Talalay, P.
  • Journal Title: Proc.Natl.Acad.Sci.USA 101 Pages: 2040-2045
• [Journal Article] Scaffolding of Keap1 to the actin cytoskeleton controls the function of Nrf2 as key regulator of cytoprotective phase 2 genes. (2004)
  • Author(s): Kang, MI, Kobayashi, A., Wakabayashi, N., Kim, S.G., Yamamoto, M.
  • Journal Title: Proc.Natl.Acad.Sci.USA 101 Pages: 2046-2051
• [Journal Article] Neurogenin3 delineates the earliest stages of spermatogenesis in the mouse testis. (2004)
  • Author(s): Yoshida, S., Takakura, A., Ohbo, K., Abe, K., Wakabayashi, J., Yamamoto, M., Suda, T., Nabeshima, Y-i.
  • Journal Title: Dev.Biol 269 Pages: 447-458
• [Journal Article] Interleukin-6 and tumor necrosis factor-a in patients with obstructive sleep apnea-hypop nea syndrome (2004)
  • Author(s): Imagawa, S., Yamaguchi, Y., Ogawa, K., Obara, N., Suzuki, N., Yamamoto, M., Nagasawa, T.
  • Journal Title: Respiration 71 Pages: 24-29
• [Journal Article] Small Maf proteins are obligate transcriptional cofactors for normal keratinocyte differentiation in the Keap1-Nrf2 regulatory pathway. (2004)
  • Author(s): Motohashi, H., Katsuoka, F., Engel, J.D., Yamamot, M.
  • Journal Title: Proc.Natl.Acad.Sci.USA 101 Pages: 6379-6384
• [Journal Article] Nrf2 deficiency improves autoimmune nephritis caused by the fas mutation lpr (2004)
  • Author(s): Morito, N., Yoh, K., Hirayama, A., Itoh, K., Nose, M., Koyama, A., Yamamoto, M.
  • Journal Title: Kidney Int. 65 Pages: 1703-1713
• [Journal Article] A constitutively active arylhydrocarbon receptor induces growth inhibition of Jurkat T cells through changes in the expression of genes related to apoptosis and cell cycle arrest. (2004)
  • Author(s): Ito, T., Tsukumo, S., Suzuki, N., Motohashi, H., Yamamoto, M., Fujii-Kuriyama, Y.
  • Journal Title: J.Biol.Chem. 279 Pages: 25204-25210
• [Journal Article] Activation of hepatic Nrf2 in vivo by acetaminophen in CD-1 Mice. (2004)
  • Author(s): Goldring, C.E.P., Kitteringham, N.R., Elsby, R., Randle, L.E., Clement, Y.N., Williams, D.P., McMahon, M., Hayes, J.D., Itoh, K., Yamamoto, M., Park, B.K.
  • Journal Title: Hepatology 39 Pages: 1267-1276
• [Journal Article] In vivo and in vitro constant expression of GATA-4 in mouse postnatal Sertoli cells. (2004)
  • Author(s): Imai, T., Kawai, Y., Tadokoro, Y., Yamamoto, M., Nishimune, Y., Yomogida K.
  • Journal Title: Mol.Cell.Endocrinol. 214 Pages: 107-115
• [Journal Article] Nrf2 deficiency causes tooth decolorization due to iron transport disorder in enamel organ. (2004)
  • Author(s): Yanagawa, T., Itoh, K., Uwayama, J., Shibata, Y., Yamaguchi, A., Sato, T., Ishii, T., Yoshida, H., Yamamoto, M.
  • Journal Title: Genes Cells 9 Pages: 641-651
• [Journal Article] Redox-regulated tumover of Nrf2 is determined by at least two separate protein domains the redox sensitive Neh2 degron and the redox-insensitive Neh6 degron. (2004)
  • Author(s): McMahon, M., Thomas, N., Itoh, K., Yamamoto, M., Haves, J.D.
  • Journal Title: J.Biol.Chem. 279 Pages: 31556-31567
• [Journal Article] The transcriptional program of antibody class switching involves the repressor Bach2. (2004)
  • Author(s): Muto, A., Tashiro, S., Nakajima, O., Hoshino, H., Takahashi, S., Sakoda, E., Ikebe, D., Yamamoto, M., Igarashi, K.
  • Journal Title: Nature 429 Pages: 556-571
• [Journal Article] The transcription factor Nrf2 protects against pulmonary fibrosis. (2004)
  • Author(s): Cho, H-Y., Reddy, S.P.M., Yamamoto, M., Kleeberger, S.R.
  • Journal Title: FASEB J. 18 Pages: 1258-1260
• [Journal Article] MafT, a new member of the small Maf protein family in zebrafish. (2004)
  • Author(s): Takagi, Y., Kobayashi, M., Li, L., Suzuki, T., Nishikawa.K., Yamamoto, M.
  • Journal Title: Biochem.Biophys.Res.Connnun. 320 Pages: 62-69
• [Journal Article] Oxidative stress sensor Keap1 functions as an adaptor for Cul3-based E3 ligase to regulate proteasomal degradation of Nrf2. (2004)
  • Author(s): Kobayashi, A., Kang, M.I., Ohkawa, H., Ohtuji, M., Zenke, Y., Chiba, T., Igarashi, K., Yamamoto, M.
  • Journal Title: Mol.Cell.Biol. 24 Pages: 7310-7319
• [Journal Article] Identification of polymorphisms in the promoter region of human NRF2 gene. (2004)
  • Author(s): Yamamoto, T., Yoh, K., Kobayashi, K., Ishii, Y., Kure, S., Koyama, A., Sekizawa, K., Motohashi, H., Yamamoto, M.
  • Journal Title: Biochem.Biophys.Res.Connnun. 321 Pages: 72-79
• [Journal Article] Do b-globin, GATA-1 or EpoR regulatory domains specifically mark erythroid progenitors in transgenic reporter mice? (2004)
  • Author(s): Suzuki, N., Imagawa, S., Noguchi C.T., Yamamoto, M.
  • Journal Title: Blood 104 Pages: 2968-2969
• [Journal Article] Multiple, distant Gata2 enhancers specify temporal and tissue-specific patterning in the developing urogenital system. (2004)
  • Author(s): Khandekar, M.J., Suzuki, N., Lewton, J., Yamamoto, M., Engel, J.D.
  • Journal Title: Mol.Cell.Biol. 24 Pages: 10263-10276
• [Journal Article] Nrf2 Is Essential for the Chemopreventive Efficacy of Oltipraz against Urinary Bladder Carcinogenesis. (2004)
  • Author(s): Iida, K., Itoh, K., Kumagai, Y., Oyasu, R., Hattori, K., Kawai, K., Shimazui, T., Akaza, H., Yamamoto, M.
  • Journal Title: Cancer Res. 64 Pages: 6124-6431
• [Journal Article] Genetic ablation of Nrf2 enhances susceptibility to cigarette smoke-induced emphysema in mice (2004)
  • Author(s): Rangasamy, T., Cho, C.Y., Thimmulappa, R.K, Zhen, L, Srisuma, S.S., Kensler, T.W., Yamamoto, M., Petrache, I., Tuder, R, M, Biswal, S.
  • Journal Title: J.Clin.Invest 114 Pages: 1248-1259
• [Journal Article] A GATA-specific inhibitor (K-7174) rescues anemia induced by IL-1b, TNF-a or L-NMMA. (2004)
  • Author(s): Imagawa, S., Nakano, Y., Obara, N., Suzuki, N., Doi, T., Kodama, T., Nagasawa, T., Yamamoto, M.
  • Journal Title: Blood 104 Pages: 4300-4307
• [Journal Article] Transcription factor Nrf2 is essential for induction of NAD(P)H : quinone oxidoreductase 1, glutathione S-transferases, and glutamate cysteine ligase by broccoli seeds and isothiocyanates. (2004)
  • Author(s): McWalter, G.K., Higgins, L.G., McLellan, L.I., Henderson, C.J., Song, L., Thomalley, P.J., Itoh, K., Yamamoto, M., Hayes, J.D.
  • Journal Title: J.Nutr. 134 Pages: 3499S-3506S
• [Journal Article] Leukemogenesis caused by incapacitated GATA-1 function. (2004)
  • Author(s): Shimizu, R., Kuroha, T., Ohneda, 0., Pan, X., Ohneda, K., Takahashi, S., Philipsen, S., Yamamoto, M.
  • Journal Title: Mol.Cell.Biol. 24 Pages: 10814-10825
• [Journal Article] Gene expression profiling of NRF2-mediated protection against oxidative injury. (2004)
  • Author(s): Cho, H-Y., Reddy, S.P., DeBiase, A., Yamamoto, M, Kleeberger, S.R.
  • Journal Title: Free Rad.Biol.Med. 38 Pages: 325-343
• [Journal Article] An activator protein-1-Notch-4 angiogenic vascular remodeling pathway. (2004)
  • Author(s): Wu, J., Iwata, F., Grass, J.A., Elnitski, L., Ohneda, O., Yamamoto, M., Bresnick, E.H.
  • Journal Title: Mol.Cell.Biol. 25 Pages: 1458-1474
• [Journal Article] Unique function of the Nrf2-Keap1 pathway in the inducible expression of antioxidant and detoxifying enzymes. (2004)
  • Author(s): Kobayashi, A., Ohta, T, Yamamoto, M.
  • Journal Title: Methods Enzymol. 378 Pages: 1208-1213
• [Journal Article] Molecular mechanism activating Nrf2-Keap1 pathway in regulation of adaptive response to electrophiles. (2004)
  • Author(s): Itoh, K., Tong, K.I., Yamamoto, M.
  • Journal Title: Free Radical Biol.Med. 36 Pages: 1208-1121
• [Journal Article] Nrf2-Keap1 defines a physiologically important stress response mechanism. (2004)
  • Author(s): Motohashi, H, Yamamoto, M.
  • Journal Title: Trends Mal.Med. 10 Pages: 549-557
• [Journal Article] GATA転写因子からみる赤血球の発生・分化 (2004)
  • Author(s): 長野真澄, 山本雅之
  • Journal Title: 実験医学 22 Pages: 361-366
• [Journal Article] 親電子性物質応答の分子機構 (2004)
  • Author(s): 伊東 健, 山本雅之
  • Journal Title: 生化学 76 Pages: 339-348
• [Journal Article] Keap1-Nrf2システムによる抗酸化酵素群の統一的発現制御機構 (2004)
  • Author(s): 細谷朋方, 山本雅之
  • Journal Title: 分子呼吸器病 8 Pages: 147-150
• [Journal Article] 血色素の分子進化 (2004)
  • Author(s): 鈴木教郎, 山本雅之
  • Journal Title: 日本医事新報 4177 Pages: 99-100
• [Journal Article] 転写因子と血液疾患 (2004)
  • Author(s): 峯岸直子, 山本雅之
  • Journal Title: 臨床検査 48 Pages: 473-781
• [Journal Article] GATA-1ノックダウンマウス (2004)
  • Author(s): 清水律子, 山本雅之
  • Journal Title: Mebio 21 Pages: 58-62
• [Journal Article] GATA-1変異と白血病-転写因子の質的異常と量的異常 (2004)
  • Author(s): 長野真澄, 清水律子, 長野真澄, 清水律子,
  • Journal Title: 医学の歩み 211 Pages: 159-162
• [Journal Article] Transgenic expression of Bach1 transcription factor results in megakaryocytic impairment.
  • Author(s): Toki, T., Katsuoka, F., Kanezaki, R., Xu, G., Kurotaki, H., Sun, J., Kamio, T., Watanabe, S., Tandai, S., Terui, K., Yagihashi, S., Komatsu, N., Igarashi, K, Yamamoto, M, Ito, E.
  • Journal Title: Blood (印刷中)